Anti-Aquaporin 4 [D15107]

Name: Anti-Aquaporin 4 [D15107]
SKU: Ab03048
Availability: InStock

Description:

Recombinant monoclonal antibody to Aquaporin 4. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma D15107.

Clone Name:

D15107

Published Application(s):

inhibit, IP, ELISA, FC

Published Species Reactivity:

Human, Mouse

Products based on this clone:

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Ab03048-23.0 Anti-Aquaporin 4 [D15107]
Human, Mouse
Rabbit IgG
Purified
In Stock

Ab03048-2.0 Anti-Aquaporin 4 [D15107]
Human, Mouse
Mouse IgG2a
Purified
In Stock

Ab03048-2.3 Anti-Aquaporin 4 [D15107]
Human, Mouse
Mouse IgG2a
Fc Silent™
Purified
Ships in 4-5 weeks

Ab03048-10.0 Anti-Aquaporin 4 [D15107]
Human, Mouse
Human IgG1
Purified
In Stock

Ab03048-10.3 Anti-Aquaporin 4 [D15107]
Human, Mouse
Human IgG1
Fc Silent™
Purified
Ships in 4-5 weeks

UniProt Accession Number of Target Protein: P55087

Alternative Name(s) of Target: AQP4; AQP-4; Aquaporin-4; MIWC; WCH4; Mercurial-insensitive water channel
Immunogen: The original antibody was generated by immunizing an aquaporin 4 knock out mice with human AQP4 as an immunogen expressed on the surface of baculovirus.

Specificity: This antibody recognizes and binds the extracellular domains of both the M1 and M23 isoforms of human aquaporin 4 (AQP4). It also binds the M23 isoform of mouse aquaporin 4, and does not recognize the M1 isoform of mouse aquaporin 4. AQP4 is a water channel expressed on astrocytes throughout the CNS and in skeletal muscle and some epithelial cells outside of the CNS. It plays an important role in brain water homeostasis and in glymphatic solute transport. Neuromyelitis optica spectrum disorder (NMOSD) is an inflammatory disease of the central nervous system (CNS) that is associated with demyelination and neurological deficits in optic nerve, spinal cord and brainstem. Most NMOSD patients are seropositive for IgG autoantibodies (NMO-IgG) directed against extracellular epitopes of aquaporin-4 (AQP4). Prevention of the binding of NMO-IgG, therefore, may alleviate the disease.

Application Notes: This antibody binds extracellular domain of human aquaporin 4 with a very high binding affinity. The binding of this antibody inhibits the binding of NMO-IgG to AQP4 (PMID: 23746426). The binding affinity of this antibody for human AQP4 M23 isoform expressed on the surface of baculovirus and CHO cells was tested with ELISA (PMID: 26398585; WO2014196658). The binding characterization of this antibody was done using flow cytometry. This antibody can be used for the immunoprecipitation of human AQP4 but not mouse AQP4 (WO2014196658; PMID: 23909421). This antibody is not suitable for detection of denatured human AQP4 in a western blot assay (WO2014196658). Live imaging using Alexa‐Fluor‐555‐labelled antibodies revealed that this antibody more rapidly bound to cells expressing human aquaporin‐4 and strongly enhanced endocytosis and lysosomal degradation of AQP4, especially in cells expressing the M1 isoform alone. Chimeric D15107 prevented complement‐dependent cytotoxicity induced by NMO‐IgG from patient sera in vitro (PMID: 26398585).

Antibody first published in: Miyazaki et al. Establishment of monoclonal antibodies against the extracellular domain that block binding of NMO-IgG to AQP. J Neuroimmunol. 2013 Jul 15;260(1-2):107-16. PMID:23746426
Note on publication: Describes the generation of antibodies against extracellular domain of AQP4 and explores the therapeutic application of this antibody in the treatment of neuromyelitis optica patients.

Immunofluorescence staining of A431 cells using Ab03048-23.0. Click on image for more details.

Immunofluorescence staining of A431 cells with anti-Aquaporin 4 antibody D15107 (Ab03048). Immunofluorescence analysis of paraformaldehyde fixed A431 cells on Shi-fix™ coverslips stained with anti-Aquaporin 4 antibody D15107 (Ab03048-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, Ab03048-23.0, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.

Western blot of unboiled and boiled mouse skeletal muscle tissue lysates using Ab03048-23.0. Click on image for more details.

Western blot using anti-Aquaporin 4 antibody D15107 (Ab03048). Unboiled (A) and boiled (B) mouse skeletal muscle tissue lysates (35µg protein in RIPA buffer) were resolved via SDS-PAGE, and the subsequent blots were probed with the chimeric rabbit version of D15107 (Ab03048-23.0) at 1µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1 hour was used, and proteins were detected by chemiluminescence.

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